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Published in: Malaria Journal 1/2021

01-12-2021 | Plasmodium Falciparum | Research

Direct contact between Plasmodium falciparum and human B-cells in a novel co-culture increases parasite growth and affects B-cell growth

Authors: Sreenivasulu B. Reddy, Noemi Nagy, Caroline Rönnberg, Francesca Chiodi, Allan Lugaajju, Frank Heuts, Laszlo Szekely, Mats Wahlgren, Kristina E. M. Persson

Published in: Malaria Journal | Issue 1/2021

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Abstract

Background

Plasmodium falciparum parasites cause malaria and co-exist in humans together with B-cells for long periods of time. Immunity is only achieved after repeated exposure. There has been a lack of methods to mimic the in vivo co-occurrence, where cells and parasites can be grown together for many days, and it has been difficult with long time in vitro studies.

Methods and results

A new method for growing P. falciparum in 5% CO2 with a specially formulated culture medium is described. This knowledge was used to establish the co-culture of live P. falciparum together with human B-cells in vitro for 10 days. The presence of B-cells clearly enhanced parasite growth, but less so when Transwell inserts were used (not allowing passage of cells or merozoites), showing that direct contact is advantageous. B-cells also proliferated more in presence of parasites. Symbiotic parasitic growth was verified using CESS cell-line and it showed similar results, indicating that B-cells are indeed the cells responsible for the effect. In malaria endemic areas, people often have increased levels of atypical memory B-cells in the blood, and in this assay it was demonstrated that when parasites were present there was an increase in the proportion of CD19 + CD20 + CD27 − FCRL4 + B-cells, and a contraction of classical memory B-cells. This effect was most clearly seen when direct contact between B-cells and parasites was allowed.

Conclusions

These results demonstrate that P. falciparum and B-cells undoubtedly can affect each other when allowed to multiply together, which is valuable information for future vaccine studies.
Literature
1.
go back to reference Murray CJL, Rosenfeld LC, Lim SS, Andrews KG, Foreman KJ, Haring D, et al. Global malaria mortality between 1980 and 2010: a systematic analysis. Lancet. 2012;379:413–31.CrossRefPubMed Murray CJL, Rosenfeld LC, Lim SS, Andrews KG, Foreman KJ, Haring D, et al. Global malaria mortality between 1980 and 2010: a systematic analysis. Lancet. 2012;379:413–31.CrossRefPubMed
2.
go back to reference Beeson JG, Osier FHA, Engwerda CR. Recent insights into humoral and cellular immune responses against malaria. Trends Parasitol. 2008;24:578–84.CrossRefPubMed Beeson JG, Osier FHA, Engwerda CR. Recent insights into humoral and cellular immune responses against malaria. Trends Parasitol. 2008;24:578–84.CrossRefPubMed
3.
go back to reference Dorfman JR, Bejon P, Ndungu FM, Langhorne J, Kortok MM, Lowe BS, et al. B-cell memory to 3 Plasmodium falciparum blood-stage antigens in a malaria-endemic area. J Infect Dis. 2005;191:1623–30.CrossRefPubMed Dorfman JR, Bejon P, Ndungu FM, Langhorne J, Kortok MM, Lowe BS, et al. B-cell memory to 3 Plasmodium falciparum blood-stage antigens in a malaria-endemic area. J Infect Dis. 2005;191:1623–30.CrossRefPubMed
4.
go back to reference Langhorne J, Ndungu FM, Sponaas A-M, Marsh K. Immunity to malaria: more questions than answers. Nat Immunol. 2008;9:725–32.CrossRefPubMed Langhorne J, Ndungu FM, Sponaas A-M, Marsh K. Immunity to malaria: more questions than answers. Nat Immunol. 2008;9:725–32.CrossRefPubMed
5.
6.
go back to reference Marsh K, Otoo L, Hayes RJ, Carson DC, Greenwood BM. Antibodies to blood stage antigens of Plasmodium falciparum in rural Gambians and their relation to protection against infection. Trans R Soc Trop Med Hyg. 1989;83:293–303.CrossRefPubMed Marsh K, Otoo L, Hayes RJ, Carson DC, Greenwood BM. Antibodies to blood stage antigens of Plasmodium falciparum in rural Gambians and their relation to protection against infection. Trans R Soc Trop Med Hyg. 1989;83:293–303.CrossRefPubMed
7.
go back to reference Giha HA, Staalsoe T, Dodoo D, Elhassan IM, Roper C, Satti GMH, et al. Nine-year longitudinal study of antibodies to variant antigens on the surface of Plasmodium falciparum-infected erythrocytes. Infect Immun. 1999;67:4092–8.CrossRefPubMedPubMedCentral Giha HA, Staalsoe T, Dodoo D, Elhassan IM, Roper C, Satti GMH, et al. Nine-year longitudinal study of antibodies to variant antigens on the surface of Plasmodium falciparum-infected erythrocytes. Infect Immun. 1999;67:4092–8.CrossRefPubMedPubMedCentral
8.
go back to reference Cavanagh DR, Elhassan IM, Roper C, Robinson VJ, Giha H, Holder AA, et al. A longitudinal study of type-specific antibody responses to Plasmodium falciparum merozoite surface protein-1 in an area of unstable malaria in Sudan. J Immunol. 1998;161:347–59.PubMed Cavanagh DR, Elhassan IM, Roper C, Robinson VJ, Giha H, Holder AA, et al. A longitudinal study of type-specific antibody responses to Plasmodium falciparum merozoite surface protein-1 in an area of unstable malaria in Sudan. J Immunol. 1998;161:347–59.PubMed
9.
go back to reference Gupta S, Snow RW, Donnelly CA, Marsh K, Newbold C. Immunity to non-cerebral severe malaria is acquired after one or two infections. Nat Med. 1999;5:340–3.CrossRefPubMed Gupta S, Snow RW, Donnelly CA, Marsh K, Newbold C. Immunity to non-cerebral severe malaria is acquired after one or two infections. Nat Med. 1999;5:340–3.CrossRefPubMed
10.
go back to reference Weidanz WP, Long CA. The role of T cells in immunity to malaria. Progr Allergy. 1988;41:215–52. Weidanz WP, Long CA. The role of T cells in immunity to malaria. Progr Allergy. 1988;41:215–52.
11.
go back to reference Cohen S, McGregor IA, Carrington S. Gamma-globulin and acquired immunity to human malaria. Nature. 1961;192:733–7.CrossRefPubMed Cohen S, McGregor IA, Carrington S. Gamma-globulin and acquired immunity to human malaria. Nature. 1961;192:733–7.CrossRefPubMed
12.
go back to reference Wipasa J, Elliott S, Xu H, Good MF. Immunity to asexual blood stage malaria and vaccine approaches. Immunol Cell Biol. 2002;80:401–14.CrossRefPubMed Wipasa J, Elliott S, Xu H, Good MF. Immunity to asexual blood stage malaria and vaccine approaches. Immunol Cell Biol. 2002;80:401–14.CrossRefPubMed
14.
go back to reference Weiss GE, Crompton PD, Li S, Walsh LA, Moir S, Traore B, et al. Atypical memory B-cells are greatly expanded in individuals living in a malaria-endemic area. J Immunol. 2009;183:2176–82.CrossRefPubMed Weiss GE, Crompton PD, Li S, Walsh LA, Moir S, Traore B, et al. Atypical memory B-cells are greatly expanded in individuals living in a malaria-endemic area. J Immunol. 2009;183:2176–82.CrossRefPubMed
15.
go back to reference Weiss GE, Traore B, Kayentao K, Ongoiba A, Doumbo S, Doumtabe D, et al. The Plasmodium falciparum-specific human memory B-cell compartment expands gradually with repeated malaria infections. PLoS Pathog. 2010;6: e1000912.CrossRefPubMedPubMedCentral Weiss GE, Traore B, Kayentao K, Ongoiba A, Doumbo S, Doumtabe D, et al. The Plasmodium falciparum-specific human memory B-cell compartment expands gradually with repeated malaria infections. PLoS Pathog. 2010;6: e1000912.CrossRefPubMedPubMedCentral
16.
go back to reference Illingworth J, Butler NS, Roetynck S, Mwacharo J, Pierce SK, Bejon P, et al. Chronic exposure to Plasmodium falciparum is associated with phenotypic evidence of B and T Cell exhaustion. J Immunol. 2013;190:1038–47.CrossRefPubMed Illingworth J, Butler NS, Roetynck S, Mwacharo J, Pierce SK, Bejon P, et al. Chronic exposure to Plasmodium falciparum is associated with phenotypic evidence of B and T Cell exhaustion. J Immunol. 2013;190:1038–47.CrossRefPubMed
17.
go back to reference Weiss GE, Clark EH, Li S, Traore B, Kayentao K, Ongoiba A, et al. A positive correlation between atypical memory B-cells and Plasmodium falciparum transmission intensity in cross-sectional studies in Peru and Mali. PLoS ONE. 2011;6: e15983.CrossRefPubMedPubMedCentral Weiss GE, Clark EH, Li S, Traore B, Kayentao K, Ongoiba A, et al. A positive correlation between atypical memory B-cells and Plasmodium falciparum transmission intensity in cross-sectional studies in Peru and Mali. PLoS ONE. 2011;6: e15983.CrossRefPubMedPubMedCentral
18.
go back to reference Yeo KT, Embury P, Anderson T, Mungai P, Malhotra I, King C, et al. HIV, cytomegalovirus, and malaria infections during pregnancy lead to inflammation and shifts in memory B-cell subsets in Kenyan neonates. J Immunol. 2019;202:1465–78.CrossRefPubMedPubMedCentral Yeo KT, Embury P, Anderson T, Mungai P, Malhotra I, King C, et al. HIV, cytomegalovirus, and malaria infections during pregnancy lead to inflammation and shifts in memory B-cell subsets in Kenyan neonates. J Immunol. 2019;202:1465–78.CrossRefPubMedPubMedCentral
19.
go back to reference Amu S, Ruffin N, Rethi B, Chiodi F. Impairment of B-cell functions during HIV-1 infection. AIDS. 2013;27:2323–34.CrossRefPubMed Amu S, Ruffin N, Rethi B, Chiodi F. Impairment of B-cell functions during HIV-1 infection. AIDS. 2013;27:2323–34.CrossRefPubMed
20.
go back to reference Kardava L, Moir S, Wang W, Ho J, Buckner CM, Posada JG, et al. Attenuation of HIV-associated human B-cell exhaustion by siRNA downregulation of inhibitory receptors. J Clin Invest. 2011;121:2614–24.CrossRefPubMedPubMedCentral Kardava L, Moir S, Wang W, Ho J, Buckner CM, Posada JG, et al. Attenuation of HIV-associated human B-cell exhaustion by siRNA downregulation of inhibitory receptors. J Clin Invest. 2011;121:2614–24.CrossRefPubMedPubMedCentral
21.
go back to reference Moir S, Ho J, Malaspina A, Wang W, DiPoto AC, O’Shea MA, et al. Evidence for HIV-associated B-cell exhaustion in a dysfunctional memory B-cell compartment in HIV-infected viremic individuals. J Exp Med. 2008;205:1797–805.CrossRefPubMedPubMedCentral Moir S, Ho J, Malaspina A, Wang W, DiPoto AC, O’Shea MA, et al. Evidence for HIV-associated B-cell exhaustion in a dysfunctional memory B-cell compartment in HIV-infected viremic individuals. J Exp Med. 2008;205:1797–805.CrossRefPubMedPubMedCentral
22.
23.
go back to reference Ribacke U, Moll K, Albrecht L, Ismail HA, Normark J, Flaberg E, et al. Improved in vitro culture of Plasmodium falciparum permits establishment of clinical isolates with preserved multiplication, invasion and rosetting phenotypes. PLoS ONE. 2013;8: e69781.CrossRefPubMedPubMedCentral Ribacke U, Moll K, Albrecht L, Ismail HA, Normark J, Flaberg E, et al. Improved in vitro culture of Plasmodium falciparum permits establishment of clinical isolates with preserved multiplication, invasion and rosetting phenotypes. PLoS ONE. 2013;8: e69781.CrossRefPubMedPubMedCentral
24.
go back to reference Rono J, Färnert A, Olsson D, Osier F, Rooth I, Persson KEM. Plasmodium falciparum line-dependent association of in vitro growth-inhibitory activity and risk of malaria. Infect Immun. 2012;80:1900–8.CrossRefPubMedPubMedCentral Rono J, Färnert A, Olsson D, Osier F, Rooth I, Persson KEM. Plasmodium falciparum line-dependent association of in vitro growth-inhibitory activity and risk of malaria. Infect Immun. 2012;80:1900–8.CrossRefPubMedPubMedCentral
25.
go back to reference Flaberg E, Sabelström P, Strandh C, Szekely L. Extended field laser confocal microscopy (EFLCM): combining automated Gigapixel image capture with in silicovirtual microscopy. BMC Med Imag. 2008;8:13.CrossRef Flaberg E, Sabelström P, Strandh C, Szekely L. Extended field laser confocal microscopy (EFLCM): combining automated Gigapixel image capture with in silicovirtual microscopy. BMC Med Imag. 2008;8:13.CrossRef
26.
go back to reference Crompton PD, Mircetic M, Weiss G, Baughman A, Huang C-Y, Topham DJ, et al. The TLR9 ligand CpG promotes the acquisition of Plasmodium falciparum-specific memory B-cells in malaria-naïve individuals. J Immunol. 2009;182:3318–26.CrossRefPubMed Crompton PD, Mircetic M, Weiss G, Baughman A, Huang C-Y, Topham DJ, et al. The TLR9 ligand CpG promotes the acquisition of Plasmodium falciparum-specific memory B-cells in malaria-naïve individuals. J Immunol. 2009;182:3318–26.CrossRefPubMed
27.
go back to reference Dobaño C, Santano R, Vidal M, Jiménez A, Jairoce C, Ubillos I, et al. Differential patterns of IgG subclass responses to Plasmodium falciparum antigens in relation to malaria protection and RTS,S vaccination. Front Immunol. 2019;10:439.CrossRefPubMedPubMedCentral Dobaño C, Santano R, Vidal M, Jiménez A, Jairoce C, Ubillos I, et al. Differential patterns of IgG subclass responses to Plasmodium falciparum antigens in relation to malaria protection and RTS,S vaccination. Front Immunol. 2019;10:439.CrossRefPubMedPubMedCentral
28.
go back to reference Jogdand GM, Sengupta S, Bhattacharya G, Singh SK, Barik PK, Devadas S. Inducible costimulator expressing T cells promote parasitic growth during blood stage Plasmodium berghei ANKA infection. Front Immunol. 2018;9:1041.CrossRefPubMedPubMedCentral Jogdand GM, Sengupta S, Bhattacharya G, Singh SK, Barik PK, Devadas S. Inducible costimulator expressing T cells promote parasitic growth during blood stage Plasmodium berghei ANKA infection. Front Immunol. 2018;9:1041.CrossRefPubMedPubMedCentral
29.
go back to reference Struik SS, Omer FM, Artavanis-Tsakonas K, Riley EM. Uninfected erythrocytes inhibit Plasmodium falciparum-induced cellular immune responses in whole-blood assays. Blood. 2004;103:3084–92.CrossRefPubMed Struik SS, Omer FM, Artavanis-Tsakonas K, Riley EM. Uninfected erythrocytes inhibit Plasmodium falciparum-induced cellular immune responses in whole-blood assays. Blood. 2004;103:3084–92.CrossRefPubMed
30.
go back to reference Donati D, Zhang LP, Chen Q, Chêne A, Flick K, Nyström M, et al. Identification of a polyclonal B-cell activator in Plasmodium falciparum. Infect Immun. 2004;72:5412–8.CrossRefPubMedPubMedCentral Donati D, Zhang LP, Chen Q, Chêne A, Flick K, Nyström M, et al. Identification of a polyclonal B-cell activator in Plasmodium falciparum. Infect Immun. 2004;72:5412–8.CrossRefPubMedPubMedCentral
31.
go back to reference Donati D, Mok B, Chêne A, Xu H, Thangarajh M, Glas R, et al. Increased B-cell survival and preferential activation of the memory compartment by a malaria polyclonal B-cell activator. J Immunol. 2006;177:3035–44.CrossRefPubMed Donati D, Mok B, Chêne A, Xu H, Thangarajh M, Glas R, et al. Increased B-cell survival and preferential activation of the memory compartment by a malaria polyclonal B-cell activator. J Immunol. 2006;177:3035–44.CrossRefPubMed
32.
go back to reference Derkach A, Otim I, Pfeiffer RM, Onabajo OO, Legason ID, Nabalende H, et al. Associations between IgG reactivity to Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) antigens and Burkitt lymphoma in Ghana and Uganda case-control studies. EBioMedicine. 2019;39:358–68.CrossRefPubMed Derkach A, Otim I, Pfeiffer RM, Onabajo OO, Legason ID, Nabalende H, et al. Associations between IgG reactivity to Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) antigens and Burkitt lymphoma in Ghana and Uganda case-control studies. EBioMedicine. 2019;39:358–68.CrossRefPubMed
33.
go back to reference Muellenbeck MF, Ueberheide B, Amulic B, Epp A, Fenyo D, Busse CE, et al. Atypical and classical memory B-cells produce Plasmodium falciparum neutralizing antibodies. J Exp Med. 2013;210:389–99.CrossRefPubMedPubMedCentral Muellenbeck MF, Ueberheide B, Amulic B, Epp A, Fenyo D, Busse CE, et al. Atypical and classical memory B-cells produce Plasmodium falciparum neutralizing antibodies. J Exp Med. 2013;210:389–99.CrossRefPubMedPubMedCentral
34.
go back to reference Murugan R, Buchauer L, Triller G, Kreschel C, Costa G, Pidelaserra Martí G, et al. Clonal selection drives protective memory B-cell responses in controlled human malaria infection. Sci Immunol. 2018;3: eaap8029.CrossRefPubMed Murugan R, Buchauer L, Triller G, Kreschel C, Costa G, Pidelaserra Martí G, et al. Clonal selection drives protective memory B-cell responses in controlled human malaria infection. Sci Immunol. 2018;3: eaap8029.CrossRefPubMed
35.
go back to reference Capolunghi F, Cascioli S, Giorda E, Rosado MM, Plebani A, Auriti C, et al. CpG drives human transitional B-cells to terminal differentiation and production of natural antibodies. J Immunol. 2008;180:800–8.CrossRefPubMed Capolunghi F, Cascioli S, Giorda E, Rosado MM, Plebani A, Auriti C, et al. CpG drives human transitional B-cells to terminal differentiation and production of natural antibodies. J Immunol. 2008;180:800–8.CrossRefPubMed
36.
go back to reference Siewe B, Nipper AJ, Sohn H, Stapleton JT, Landay A. FcRL4 expression identifies a pro-inflammatory B-cell Subset in viremic HIV-infected subjects. Frontiers Immunol. 2017;8:1339.CrossRef Siewe B, Nipper AJ, Sohn H, Stapleton JT, Landay A. FcRL4 expression identifies a pro-inflammatory B-cell Subset in viremic HIV-infected subjects. Frontiers Immunol. 2017;8:1339.CrossRef
37.
go back to reference Jourdan M, Robert N, Cren M, Thibaut C, Duperray C, Kassambara A, et al. Characterization of human FCRL4-positive B-cells. PLoS ONE. 2017;12: e0179793.CrossRefPubMedPubMedCentral Jourdan M, Robert N, Cren M, Thibaut C, Duperray C, Kassambara A, et al. Characterization of human FCRL4-positive B-cells. PLoS ONE. 2017;12: e0179793.CrossRefPubMedPubMedCentral
38.
go back to reference Lugaajju A, Reddy SB, Wahlgren M, Kironde F, Persson KEM. Development of Plasmodium falciparum specific naïve, atypical, memory and plasma B-cells during infancy and in adults in an endemic area. Malar J. 2017;16:37.CrossRefPubMedPubMedCentral Lugaajju A, Reddy SB, Wahlgren M, Kironde F, Persson KEM. Development of Plasmodium falciparum specific naïve, atypical, memory and plasma B-cells during infancy and in adults in an endemic area. Malar J. 2017;16:37.CrossRefPubMedPubMedCentral
Metadata
Title
Direct contact between Plasmodium falciparum and human B-cells in a novel co-culture increases parasite growth and affects B-cell growth
Authors
Sreenivasulu B. Reddy
Noemi Nagy
Caroline Rönnberg
Francesca Chiodi
Allan Lugaajju
Frank Heuts
Laszlo Szekely
Mats Wahlgren
Kristina E. M. Persson
Publication date
01-12-2021
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2021
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/s12936-021-03831-x

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